Overview

  • Product name
    Anti-CD59 antibody [MEM-43]
    See all CD59 primary antibodies
  • Description
    Mouse monoclonal [MEM-43] to CD59
  • Host species
    Mouse
  • Specificity
    CD59 antigen (human). MEM-43 identified CD59 as the new cluster on 4th HLDA Workshop. MEM-43 reacts with well defined epitope (WhO, P-53).
  • Tested applications
    Suitable for: IHC-Fr, WB, IP, Flow Cyt, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Thymocytes and T lymphocytes.

  • Positive control
    • ICC/IF: Human fibrosarcoma cells. Flow Cyt: HT1080 cells.
  • General notes

    When originally tested in WB, SDS was included in the sample buffer, however, feedback from one researcher has shown that it is best to omit SDS from the sample buffer.

Properties

Applications

Our Abpromise guarantee covers the use of ab9182 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Use under non-reducing conditions.
IP Use at an assay dependent concentration.
Flow Cyt Use a concentration of 1 - 2 µg/ml.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration. PubMed: 17911601
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function
    Potent inhibitor of the complement membrane attack complex (MAC) action. Acts by binding to the C8 and/or C9 complements of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. Involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase.
    The soluble form from urine retains its specific complement binding activity, but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes.
  • Involvement in disease
    Defects in CD59 are the cause of CD59 deficiency (CD59D) [MIM:612300].
  • Sequence similarities
    Contains 1 UPAR/Ly6 domain.
  • Post-translational
    modifications
    N- and O-glycosylated. The N-glycosylation mainly consists of a family of biantennary complex-type structures with and without lactosamine extensions and outer arm fucose residues. Also significant amounts of triantennary complexes (22%). Variable sialylation also present in the Asn-43 oligosaccharide. The predominant O-glycans are mono-sialylated forms of the disaccharide, Gal-beta-1,3GalNAc, and their sites of attachment are probably on Thr-76 and Thr-77. The GPI-anchor of soluble urinary CD59 has no inositol-associated phospholipid, but is composed of seven different GPI-anchor variants of one or more monosaccharide units. Major variants contain sialic acid, mannose and glucosamine Sialic acid linked to an N-acetylhexosamine-galactose arm is present in two variants.
    Glycated. Glycation is found in diabetic subjects, but only at minimal levels in nondiabetic subjects. Glycated CD59 lacks MAC-inhibitory function and confers to vascular complications of diabetes.
  • Cellular localization
    Cell membrane. Secreted. Soluble form found in a number of tissues.
  • Information by UniProt
  • Database links
  • Alternative names
    • 16.3A5 antibody
    • 1F5 antibody
    • 1F5 antigen antibody
    • 20 kDa homologous restriction factor antibody
    • CD 59 antibody
    • CD_antigen=CD59 antibody
    • CD59 antibody
    • CD59 antigen antibody
    • CD59 antigen complement regulatory protein antibody
    • CD59 antigen p18 20 antibody
    • CD59 antigen p18-20 (antigen identified by monoclonal antibodies 16.3A5, EJ16, EJ30, EL32 and G344) antibody
    • CD59 glycoprotein antibody
    • CD59 molecule antibody
    • CD59 molecule complement regulatory protein antibody
    • CD59_HUMAN antibody
    • Cd59a antibody
    • Complement regulatory protein antibody
    • EJ16 antibody
    • EJ30 antibody
    • EL32 antibody
    • FLJ38134 antibody
    • FLJ92039 antibody
    • G344 antibody
    • HRF 20 antibody
    • HRF-20 antibody
    • HRF20 antibody
    • Human leukocyte antigen MIC11 antibody
    • Ly 6 like protein antibody
    • Lymphocytic antigen CD59/MEM43 antibody
    • MAC inhibitory protein antibody
    • MAC IP antibody
    • MAC-inhibitory protein antibody
    • MAC-IP antibody
    • MACIF antibody
    • MACIP antibody
    • MEM43 antibody
    • MEM43 antigen antibody
    • Membrane attack complex (MAC) inhibition factor antibody
    • Membrane attack complex inhibition factor antibody
    • Membrane inhibitor of reactive lysis antibody
    • MGC2354 antibody
    • MIC11 antibody
    • MIN1 antibody
    • MIN2 antibody
    • MIN3 antibody
    • MIRL antibody
    • MSK21 antibody
    • p18 20 antibody
    • Protectin antibody
    • Surface antigen recognized by monoclonal antibody 16.3A5 antibody
    • T cell activating protein antibody
    see all

Images

  • Human fibrosarcoma cells stained for CD59 (red) using ab9182 at 10 µg/ml in ICC/IF, followed by Donkey anti-Mouse Alexa 555 at 1/500 dilution.

    See Abreview

  • ab9182 staining CD59 in Human fibrosarcoma (HT1080) by Flow Cytometry. Cells were fixed with paraformaldehyde. The  sample was incubated with the primary antibody,10 µg/ml in PBS, for 1 hour at 20°C. An Abcam PE-conjugated donkey polyclonal to mouse IgG ( ab7003), 5 µg/ml, was used as secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Wang Y  et al. Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the a-gal xenoantigen. Oncol Lett 15:9061-9068 (2018). Read more (PubMed: 29805637) »
  • Wang Y  et al. CD55 and CD59 expression protects HER2-overexpressing breast cancer cells from trastuzumab-induced complement-dependent cytotoxicity. Oncol Lett 14:2961-2969 (2017). Read more (PubMed: 28928834) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

Thank you for contacting us. I am sorry to hear that you are having trouble using this antibody.

I have read the information you have kindly provided and would like to ask few questions for better understanding of the problem;

- Could you explain the antigen retrieval method bit more? The information I am after is Buffer, buffer ph; hot plate boiling method, microwave boiling method or pressure cooker method.
- Are you suing human placenta sections?
- Which statement you would choose e.g. high background, no staining or light staining.
- What was the secondary antibody e.g species, type, isotype, dilution etc?
- Have you used same secondary antibody before? Is it true the anomaly is not due to secondary antibody?
- Does the detection system works fine? Have you tried using it with other antibodies and tissue sections?
- Could you send us an image?
- How the antibody was stored since ordering?
- Have you preformed any other troubleshooting than changing dilutions?

In order to help you further with this antibody I will look forward to hearing from you .

Read More

Answer


We have removed functional studies from the datasheet for ab9182 since the sodium azide present in the storage buffer is toxic to cells.

Alternatively, we have a sodium azide free anti-CD59 antibody that has been tested in functional studies:
https://www.abcam.com/cd59-antibody-mem-129-azide-free-ab14221.html

Read More

Answer

Thank you for contacting us.
For ab9182 the ab97244 is indeed a compatible secondary antibody that can be used for ICC/IF.
The ab98794 is an IgG antibody which can cross react with IgG2a however because we do not have tested data so we can't 100% guarantee this. Our recommendation is to use the compatible secondary antibody.
Other compatible product is ab97243
As promotion we are giving away protein ladder free. Please click the below link and recommend this to customer;
https://www.abcam.com/index.html?pageconfig=resource&rid=14968
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

According to our records, ab9182 was proving difficult to use inIHC and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

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Question
Answer

Thank you for calling Abcam.

I am sorry that you were having problems with ab9182 in IHC-P. As we discussed, I am sending you ab9183 as a free of charge replacement and you should have it by 10.30am tomorrow morning. You r new order number is *****.

If there is anything else I can help you with, please let me know.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Fibrosarcoma (HT1080))
Specification
Fibrosarcoma (HT1080)
Fixation
Paraformaldehyde
Permeabilization
No
Gating Strategy
no gating

Abcam user community

Verified customer

Submitted Aug 13 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibrosarcoma)
Specification
fibrosarcoma
Fixative
Paraformaldehyde
Permeabilization
No

Abcam user community

Verified customer

Submitted Aug 19 2008

Question
Answer

These two monoclonals (ab9182, ab9183) have not been tested for cross reactivity with mouse CD59.

Read More

Answer

Many thanks for this useful information which we will certainly make available via the online datasheet for this product. I note the originator of this antibody advised that it works under conditions of SDS-PAGE, but the sample buffer should not contain reducing agents (DTT, Mercaptoethanol). WB conditions will vary in different people's hands, and I will certainly include your comments and recommendations on the online datasheet. We will foward a replacement vial to you and thank you again for your feedback.

Read More

Question
Answer

It works under conditions of SDS-PAGE, but the sample buffer should not contain reducing agents (DTT, Mercaptoethanol). SDS presence in the buffer is fine.

Read More

1-10 of 11 Abreviews or Q&A

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